Construction,Characterization,and Chromosomal Mapping of a Fosmid Library of the White-Cheeked Gibbon (Nomascus leucogenys)  

作　　者：Liping Chen Jianping Ye Yan Liu Jinghuan Wang Weiting Su Fengtang Yang Wenhui Nie 

机构地区：[1]Key Laboratory of Cellular and Molecular Evolution, Kunming Institute of Zoology, Chinese Academy of Sciences, Kunming 650223, China [2]Graduate School of Chinese Academy of Sciences, Beijing 100039, China

出　　处：《Genomics, Proteomics & Bioinformatics》2007年第3期207-215,共9页基因组蛋白质组与生物信息学报（英文版）

基　　金：the National Natural Science Foundation of China（No.30270719 and 30623007）.

摘　　要：Gibbons have experienced extensive karyotype rearrangements during evolution and represent an ideal model for studying the underlying molecular mechanism of evolutionary chromosomal rearrangements. It is anticipated that the cloning and sequence characterization of evolutionary chromosomal breakpoints will provide vital insights into the molecular force that has driven such a radical karyotype reshuffle in gibbons. We constructed and characterized a high-quality fosmid li- brary of the white-cheeked gibbon （Nomascus leucogenys） containing 192,000 non- redundant clones with an average insert size of 38 kb and 2.5-fold genome coverage. By end sequencing of 100 randomly selected fosmid clones, we generated 196 se- quence tags for the library. These end-sequenced fosmid clones were then mapped onto the chromosomes of the white-cheeked gibbon by fluorescence in situ hy- bridization~ and no spurious chimeric clone was detected. BLAST search against the human genome showed a good correlation between the number of hit clones and the number of chromosomes, an indication of unbiased chromosomal distribu- tion of the fosmid library. The chromosomal distribution of the mapped clones is also consistent with the BLAST search result against human and white-cheeked gibbon genomes. The fosmid library and the mapped clones will serve as a valu- able resource for further studying gibbons＇ chromosomal rearrangements and the underlying molecular mechanism as well as for comparative genomic study in the lesser apes.Gibbons have experienced extensive karyotype rearrangements during evolution and represent an ideal model for studying the underlying molecular mechanism of evolutionary chromosomal rearrangements. It is anticipated that the cloning and sequence characterization of evolutionary chromosomal breakpoints will provide vital insights into the molecular force that has driven such a radical karyotype reshuffle in gibbons. We constructed and characterized a high-quality fosmid li- brary of the white-cheeked gibbon （Nomascus leucogenys） containing 192,000 non- redundant clones with an average insert size of 38 kb and 2.5-fold genome coverage. By end sequencing of 100 randomly selected fosmid clones, we generated 196 se- quence tags for the library. These end-sequenced fosmid clones were then mapped onto the chromosomes of the white-cheeked gibbon by fluorescence in situ hy- bridization~ and no spurious chimeric clone was detected. BLAST search against the human genome showed a good correlation between the number of hit clones and the number of chromosomes, an indication of unbiased chromosomal distribu- tion of the fosmid library. The chromosomal distribution of the mapped clones is also consistent with the BLAST search result against human and white-cheeked gibbon genomes. The fosmid library and the mapped clones will serve as a valu- able resource for further studying gibbons＇ chromosomal rearrangements and the underlying molecular mechanism as well as for comparative genomic study in the lesser apes.

关 键 词：fosmid library white-cheeked gibbon chromosomal mapping fluorescence in situ hybridization 

分 类 号：Q78[生物学—分子生物学]